Effects of C2-ceramide on the Malme-3M melanoma cell line

Won Suk Han, Jong Yeop Yoo, Sang Woong Youn, Dong Seok Kim, Kyoung Chan Park, Sook Young Kim, Kyu Han Kim

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Ceramide is implicated in the regulation of various signaling pathways leading to proliferation, differentiation or apoptotic cell death, but there have been few investigations about the effects of ceramide on the cell growth and the melanogenesis of melanocytes. In the present study, we investigated the effects of cell-permeable ceramide on Malme-3M human melanoma cell line. MTT proliferation assay showed that C2-ceramide inhibited the growth of Malme-3M cells in a dose-dependent manner. Cell cycle analysis confirmed the inhibition of DNA synthesis by a reduction in the S phase and an increase in the G0/G1 phase. Flow cytometric analysis for apoptotic cells and morphological observations indicated that the antiproliferative effect of C2-ceramide was not due to apoptosis. We next investigated the effects of C2-ceramide on the pigmentation of Malme-3M melanoma cells. The results showed that C2-ceramide induced only a slight decrease of tyrosinase activity and melanin synthesis. To investigate the ceramide signaling pathway, we studied the influence of C2-ceramide on extracellular signal-regulated kinase (ERK) and Akt activation by Western blot. We demonstrated that the amount of phosphorylated Akt was decreased by C2-ceramide, whereas ERK was activated transiently. Because of a well-known involvement of ceramide in apoptosis, we further investigated the level of caspase-3 and HSP70 after treatment of C2-ceramide. We found that the caspase-3 was not activated and the expression of HSP70 increased moderately. In conclusion, C2-ceramide inhibited the cell growth of Malme-3M cells without the induction of apoptosis. We suggest that increased HSP70 may be related to the resistance against apoptosis.

Original languageEnglish
Pages (from-to)10-19
Number of pages10
JournalJournal of Dermatological Science
Volume30
Issue number1
DOIs
StatePublished - Oct 2002

Fingerprint

Melanoma
Cells
Ceramides
Cell Line
Apoptosis
Extracellular Signal-Regulated MAP Kinases
Cell growth
Caspase 3
Growth
Cell Cycle Resting Phase
Monophenol Monooxygenase
N-acetylsphingosine
Melanocytes
Melanins
Pigmentation
G1 Phase
Cell death
S Phase
Assays
Cell Cycle

Keywords

  • Akt
  • Ceramide
  • ERK
  • HSP70
  • Malme-3M cell

Cite this

Han, Won Suk ; Yoo, Jong Yeop ; Youn, Sang Woong ; Kim, Dong Seok ; Park, Kyoung Chan ; Kim, Sook Young ; Kim, Kyu Han. / Effects of C2-ceramide on the Malme-3M melanoma cell line. In: Journal of Dermatological Science. 2002 ; Vol. 30, No. 1. pp. 10-19.
@article{a659d102e99d4e0e98fcac2324578b6d,
title = "Effects of C2-ceramide on the Malme-3M melanoma cell line",
abstract = "Ceramide is implicated in the regulation of various signaling pathways leading to proliferation, differentiation or apoptotic cell death, but there have been few investigations about the effects of ceramide on the cell growth and the melanogenesis of melanocytes. In the present study, we investigated the effects of cell-permeable ceramide on Malme-3M human melanoma cell line. MTT proliferation assay showed that C2-ceramide inhibited the growth of Malme-3M cells in a dose-dependent manner. Cell cycle analysis confirmed the inhibition of DNA synthesis by a reduction in the S phase and an increase in the G0/G1 phase. Flow cytometric analysis for apoptotic cells and morphological observations indicated that the antiproliferative effect of C2-ceramide was not due to apoptosis. We next investigated the effects of C2-ceramide on the pigmentation of Malme-3M melanoma cells. The results showed that C2-ceramide induced only a slight decrease of tyrosinase activity and melanin synthesis. To investigate the ceramide signaling pathway, we studied the influence of C2-ceramide on extracellular signal-regulated kinase (ERK) and Akt activation by Western blot. We demonstrated that the amount of phosphorylated Akt was decreased by C2-ceramide, whereas ERK was activated transiently. Because of a well-known involvement of ceramide in apoptosis, we further investigated the level of caspase-3 and HSP70 after treatment of C2-ceramide. We found that the caspase-3 was not activated and the expression of HSP70 increased moderately. In conclusion, C2-ceramide inhibited the cell growth of Malme-3M cells without the induction of apoptosis. We suggest that increased HSP70 may be related to the resistance against apoptosis.",
keywords = "Akt, Ceramide, ERK, HSP70, Malme-3M cell",
author = "Han, {Won Suk} and Yoo, {Jong Yeop} and Youn, {Sang Woong} and Kim, {Dong Seok} and Park, {Kyoung Chan} and Kim, {Sook Young} and Kim, {Kyu Han}",
year = "2002",
month = "10",
doi = "10.1016/S0923-1811(02)00044-0",
language = "English",
volume = "30",
pages = "10--19",
journal = "Journal of Dermatological Science",
issn = "0923-1811",
publisher = "Elsevier Ireland Ltd",
number = "1",

}

Effects of C2-ceramide on the Malme-3M melanoma cell line. / Han, Won Suk; Yoo, Jong Yeop; Youn, Sang Woong; Kim, Dong Seok; Park, Kyoung Chan; Kim, Sook Young; Kim, Kyu Han.

In: Journal of Dermatological Science, Vol. 30, No. 1, 10.2002, p. 10-19.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effects of C2-ceramide on the Malme-3M melanoma cell line

AU - Han, Won Suk

AU - Yoo, Jong Yeop

AU - Youn, Sang Woong

AU - Kim, Dong Seok

AU - Park, Kyoung Chan

AU - Kim, Sook Young

AU - Kim, Kyu Han

PY - 2002/10

Y1 - 2002/10

N2 - Ceramide is implicated in the regulation of various signaling pathways leading to proliferation, differentiation or apoptotic cell death, but there have been few investigations about the effects of ceramide on the cell growth and the melanogenesis of melanocytes. In the present study, we investigated the effects of cell-permeable ceramide on Malme-3M human melanoma cell line. MTT proliferation assay showed that C2-ceramide inhibited the growth of Malme-3M cells in a dose-dependent manner. Cell cycle analysis confirmed the inhibition of DNA synthesis by a reduction in the S phase and an increase in the G0/G1 phase. Flow cytometric analysis for apoptotic cells and morphological observations indicated that the antiproliferative effect of C2-ceramide was not due to apoptosis. We next investigated the effects of C2-ceramide on the pigmentation of Malme-3M melanoma cells. The results showed that C2-ceramide induced only a slight decrease of tyrosinase activity and melanin synthesis. To investigate the ceramide signaling pathway, we studied the influence of C2-ceramide on extracellular signal-regulated kinase (ERK) and Akt activation by Western blot. We demonstrated that the amount of phosphorylated Akt was decreased by C2-ceramide, whereas ERK was activated transiently. Because of a well-known involvement of ceramide in apoptosis, we further investigated the level of caspase-3 and HSP70 after treatment of C2-ceramide. We found that the caspase-3 was not activated and the expression of HSP70 increased moderately. In conclusion, C2-ceramide inhibited the cell growth of Malme-3M cells without the induction of apoptosis. We suggest that increased HSP70 may be related to the resistance against apoptosis.

AB - Ceramide is implicated in the regulation of various signaling pathways leading to proliferation, differentiation or apoptotic cell death, but there have been few investigations about the effects of ceramide on the cell growth and the melanogenesis of melanocytes. In the present study, we investigated the effects of cell-permeable ceramide on Malme-3M human melanoma cell line. MTT proliferation assay showed that C2-ceramide inhibited the growth of Malme-3M cells in a dose-dependent manner. Cell cycle analysis confirmed the inhibition of DNA synthesis by a reduction in the S phase and an increase in the G0/G1 phase. Flow cytometric analysis for apoptotic cells and morphological observations indicated that the antiproliferative effect of C2-ceramide was not due to apoptosis. We next investigated the effects of C2-ceramide on the pigmentation of Malme-3M melanoma cells. The results showed that C2-ceramide induced only a slight decrease of tyrosinase activity and melanin synthesis. To investigate the ceramide signaling pathway, we studied the influence of C2-ceramide on extracellular signal-regulated kinase (ERK) and Akt activation by Western blot. We demonstrated that the amount of phosphorylated Akt was decreased by C2-ceramide, whereas ERK was activated transiently. Because of a well-known involvement of ceramide in apoptosis, we further investigated the level of caspase-3 and HSP70 after treatment of C2-ceramide. We found that the caspase-3 was not activated and the expression of HSP70 increased moderately. In conclusion, C2-ceramide inhibited the cell growth of Malme-3M cells without the induction of apoptosis. We suggest that increased HSP70 may be related to the resistance against apoptosis.

KW - Akt

KW - Ceramide

KW - ERK

KW - HSP70

KW - Malme-3M cell

UR - http://www.scopus.com/inward/record.url?scp=0036803330&partnerID=8YFLogxK

U2 - 10.1016/S0923-1811(02)00044-0

DO - 10.1016/S0923-1811(02)00044-0

M3 - Article

C2 - 12354415

AN - SCOPUS:0036803330

VL - 30

SP - 10

EP - 19

JO - Journal of Dermatological Science

JF - Journal of Dermatological Science

SN - 0923-1811

IS - 1

ER -